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c-kit和KITL在成年人类卵巢中的表达与定位

Expression and localisation of c-kit and KITL in the adult human ovary.

作者信息

Tuck Astrud R, Robker Rebecca L, Norman Robert J, Tilley Wayne D, Hickey Theresa E

机构信息

Robinson Research Institute, School of Paediatrics and Reproductive Health, Adelaide, South Australia, Australia.

Dame Roma Mitchell Cancer Research Laboratories, School of Medicine; University of Adelaide, Adelaide, South Australia, Australia.

出版信息

J Ovarian Res. 2015 May 26;8:31. doi: 10.1186/s13048-015-0159-x.

Abstract

BACKGROUND

The c-kit/kit ligand (KITL) signalling axis is an essential component of ovarian folliculogenesis in mammals, but little is known about expression and localisation of its key components in the ovaries of reproductive age women. This study aimed to characterise mRNA expression of c-kit and KITL isoforms and the localisation of c-kit and KITL proteins in adult human premenopausal ovaries.

METHODS

This study utilised granulosa cells obtained from the preovulatory follicles of women undergoing assisted reproduction, pieces of ovarian tissue obtained from premenopausal women undergoing gynaecological surgeries and archival paraffin-embedded premenopausal ovarian tissues. Methodology included PCR for gene expression and Western blot or immunohistochemistry for protein expression.

RESULTS

Both c-kit mRNA isoforms, known as GNNK+ and GNNK-, were detected in human ovarian cortex, while KITL protein isoforms (KITL1 and KITL2) were present in ovarian cortex and human granulosa cells. Immunohistochemistry showed expression of KITL and c-kit protein in multiple cell types within follicles throughout development, from primordial follicles to large antral follicles, in addition to atretic follicles. Oocytes of all follicle stages expressed c-kit protein exclusively. Interestingly, unlike animal models, expression of both proteins displayed a less cell-type specific distribution with immunostaining present in granulosa, theca and stromal cells, suggesting that autocrine signalling occurs within the human ovary.

CONCLUSION

The results of this study indicate that c-kit/KITL signalling also occurs in the human ovary, as established in various animal models, and may involve previously unknown autocrine signalling.

摘要

背景

c-kit/kit配体(KITL)信号轴是哺乳动物卵巢卵泡发生的重要组成部分,但对于其关键成分在育龄女性卵巢中的表达和定位了解甚少。本研究旨在表征c-kit和KITL异构体的mRNA表达以及c-kit和KITL蛋白在成年人类绝经前卵巢中的定位。

方法

本研究利用了从接受辅助生殖的女性的排卵前卵泡中获得的颗粒细胞、从接受妇科手术的绝经前女性获得的卵巢组织块以及存档的绝经前卵巢石蜡包埋组织。方法包括用于基因表达的PCR和用于蛋白质表达的蛋白质印迹或免疫组织化学。

结果

在人类卵巢皮质中检测到两种c-kit mRNA异构体,即GNNK+和GNNK-,而KITL蛋白异构体(KITL1和KITL2)存在于卵巢皮质和人类颗粒细胞中。免疫组织化学显示,从原始卵泡到大型窦状卵泡,以及闭锁卵泡,在整个卵泡发育过程中的多种细胞类型中均有KITL和c-kit蛋白的表达。所有卵泡阶段的卵母细胞均仅表达c-kit蛋白。有趣的是,与动物模型不同,这两种蛋白的表达在颗粒细胞、卵泡膜细胞和基质细胞中均有免疫染色,显示出较少的细胞类型特异性分布,这表明人类卵巢内存在自分泌信号。

结论

本研究结果表明,c-kit/KITL信号在人类卵巢中也如在各种动物模型中一样存在,并且可能涉及以前未知的自分泌信号。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/99a0/4460643/0167de450af0/13048_2015_159_Fig1_HTML.jpg

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